J. Lamotte

Kinetic and structural optimization to catalysis at low temperatures in a psychrophilic cellulase from the Antarctic bacterium Pseudoalteromonas haloplanktis

The cold-adapted cellulase CelG has been purified from the culture supernatant of the Antarctic bacterium Pseudoalteromonas haloplanktis and the gene coding for this enzyme has been cloned, sequenced and expressed in Escherichia coli. This cellulase is composed of three structurally and functionally distinct regions: an N-terminal catalytic domain belonging to glycosidase family 5 and a C-terminal cellulose-binding domain belonging to carbohydrate-binding module family 5. The linker of 107 residues connecting both domains is one of the longest found in cellulases, and optimizes substrate accessibility to the catalytic domain by drastically increasing the surface of cellulose available to a bound enzyme molecule. The psychrophilic enzyme is closely related to the cellulase Cel5 from Erwinia chrysanthemi. Both kcat and kcat/K(m) values at 4 degrees C for the psychrophilic cellulase are similar to the values for Cel5 at 30-35 degrees C, suggesting temperature adaptation of the kinetic parameters. The thermodynamic parameters of activation of CelG suggest a heat-labile, relatively disordered active site with low substrate affinity, in agreement with the experimental data. The structure of CelG has been constructed by homology modelling with a molecule of cellotetraose docked into the active site. No structural alteration related to cold-activity can be found in the catalytic cleft, whereas several structural factors in the overall structure can explain the weak thermal stability, suggesting that the loss of stability provides the required active-site mobility at low temperatures.

Conformational analysis of β and γ-lactam antibiotics

Abstract Geometry optimization, superimposition searches and conformational analysis carried on several lactam antibiotics show that reactivity with the active-site serine penicillin-binding proteins is related to a particular spatial disposition of 2 flanking functional groups — namely a CO or COH on 1 side and a carboxylate on the other — with respect to the central scissile amide bond. Such a binding entity is found in one of the most stable conformers of the tripeptide diacetyl- l -Lys- d -Ala- d -Ala conferring substrate activity, and in benzylpenicillin, cephapyrin, thienamycin, γ-lactam, the 6-spiro-epoxypenicillin S and in one epimer of lactivicin, conferring inactivating potency. This binding entity generates a particular electronic distribution and the fact that it is conserved in compounds belonging to very different chemical families strongly suggests that it is an important feature required for enzyme recognition.

Isolation and structure of sungucine: a new type of bisindoline alkaloid

Abstract From the roots of Strychnos icaja Baill. an unsymmetrical dimeric alkaloid has been isolated and called sungucine. Its original structure has been established by X-Ray diffraction; UV, IR, MS and NMR data are also given.

Synthese et structure de L'o-carboxybenzeneselenonate d'ammonium monohydrate

Abstract Oxidation of benzisothiazole by hydrogen peroxide leads to saccharine, but the same reaction with benzisoselenazole does not lead to selenosaccharine. The product of this oxidation is the ammonium o -carboxybenzeneselenonate monohydrate. Crystals are triclinic, P1, with a =6.243, b =7.598 and c =12.457 A, α =142.38, β = 104.98 and γ = 85.47°, and Z = 2. The refinement led to a final R-value of 0.056. The ammonium ion and water molecule coordinations are discussed in terms of hydrogen bonds. The coordination of the selenium atom is also described.